Based of the study on the impact of protoplasts under different conditions of enzymolysis solution, culture time, enzymolysis time and enzymolysis temperature in sequence using orthogonal test. The best optimal condition of producing protoplast was :0.5% helicase add 0.5% lysozyme and 1.0% cellulase, enzymolysis time was 3 hours , enzymolysis temperature was 32℃,under these condition the production of protoplast can be 4.25?10~(6 )per mL. Ca~(2+) and PEG can increase the regeneration ratio of protoplast.After induced mutation by complex factor under DES,UV and LiCl,the mutagenesis strain which own increasing enzyme activity and transmissibility steaty was obtained.

In our university,the training for geriatric postgraduates of clinical medicine is now in the initial stage.Therefore,it is important to explore the training model to improve the quality of postgraduate.The program focused on cultivating geriatric postgraduates' ability of diagnosis and treatment,helping them have the concept of integral service and enhancing their teaching and thinking ability of clinical sciences to provide high-level comprehensive geriatric doctors for the society.

Ras is a kind of protein closely related to neoplasms,the persistent activation of which would induce tumorigenesis and progression.RASAL1 is a Ca~(2+) regulated ras GTPase-activating-like protein, the loss or decreased expression of RASAL1 induces ras activation, then results in tumongenesis and progression.In this review we sum up the mechanism of ras activation, the relationship between neoplasms and RASAL1 and the mechanism of RASAL1 expression decreasing.

As an important device of modern medical treatment,medical monitor is a combination of modern electronics technology,computer technology and medical technology.In this paper,the development of medical monitors in China is summarized,especially the main technique characteristics of medical monitor in Chinese market.The examples of monitors made in China are presented.

Objective To explore effect of miR‐302a on proliferation and apoptosis in folate deficiency mouse embryonic stem cell (mESC) .Methods The cases were divided into complete culture medium group (control group) ,folate‐deficient culture medi‐um group (folate‐deficient group) ,folate‐deficient culture medium plus miR‐302a mimic group (miR‐302a group) .The expression of miR‐302a was examed by RT‐PCR in control group and folate‐deficient group .To construct miR‐302a mimic and then was trans‐fected into the folate‐deficient culture medium mESC .Effect of miR‐302a mimic on mESC viability was detected by MTT assay ,the effect of miR‐302a mimic on mESC apoptosis was examed by Annexin V‐FITC/PI flow dual‐staining method ,the effect of miR‐302a mimic on mESC cycle was examed by flow cytometry .The activation of phosphatidylinositol 3 kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) and expression of CyclinD1 ,p21 and p27 was assayed by Western blot . Results The expression of miR‐302a was lower in folate‐deficient group(P<0 .01) .Compared with control group ,the viability of mESC was lower ,the apoptosis of mESC was higher ,the cell cycle was arrested in G1 phase ,the level of phosphorylation of AKT and mTOR was lower ,the expression of CyclinD1 was lower ,the expression of p21 and p27 was higher in folate‐deficient group , with statistical significance (P<0 .01) .Compared with folate‐deficient group ,the viability of mESC was higher ,the apoptosis of mESC was lower ,G1 phase was shortened ,the level of phosphorylation of AKT and mTOR was higher ,the expression of CyclinD1 was higher ,the expression of p21 and p27 was lower in miR‐302a group with statistical significance (P<0 .01) .Conclusion These results suggested miR‐302a exerted anti‐apoptosis and promote cell proliferation in folate‐deficient culture medium ,which might be related to PI3K/AKT/mTOR signal pathway .

Objective To investigate the clinical effect of insulin on gestational diabetes mellitus(GDM).Methods GDM patients were randomly divided into group A and group B,group B were treated with short term intensive insulin pump therapy,and group A were treated with insulin pump in short term combined with biosynthetic human insulin therapy.Results The body mass index and diabetes family history rates pre-pregnancy in group B were significantly higher than group A(P<0.05).Logistic analysis showed that family history of BMI pre-pregnancy and diabetes family history were independent risk factors for GDM drug treatment(P<0.05).Compared with the group B,the improvement of blood glucose levels and the incidence of complications in patients with group A were better than the former(P<0.05).Conclusion Rational application of insulin can improve the efficacy of clinical treatment of GDM, reduce the incidence of complications, with high safety.

BACKGROUND:It is confirmed that the onset of osteoarthritis is closely related to inflammatory cytokines. OBJECTIVE: To investigate the levels of interleukin-6 and matrix metaloproteinase-3 in the synovial fluid of patients with osteoarthritis and their correlation with the severity of osteoarthritis. METHODS: Synovial fluid specimens of 78 patients were harvested to detect the levels of interleukin-6 and matrix metaloproteinase-3 using ELISA assay. The severity of osteoarthritis was estimated by the Hospital for Special Surgery (HSS) score. Pearson correlation analysis was adopted to explore the pairwise correlations between interleukin-6, matrix metaloproteinase-3 and HSS score. RESULTS AND CONCLUSION:The levels of interleukin-6 and matrix metaloproteinase-3 in the synovial fluid were (13.1±5.7) and (989.3±429.5) μg/L, respectively; the HSS score was (56.4±12.0) points. Pearson correlation analysis showed that the expression levels of interleukin-6 and matrix metaloproteinase-3 had a significant positive correlation (r=0.70,P< 0.001); the levels of interleukin-6 and matrix metaloproteinase-3 had a negative correlation with the HSS scores (r=-0.70,r=-0.75,P < 0.001). Experimental findings show that there is a positive correlation between interleukin-6 and matrix metaloproteinase-3 in the synovial fluids, and these two cytokines also have a positive correlation with the severity of osteoarthritis, indicating that to measure interleukin-6 and matrix metaloproteinase-3 levels in the synovial fluids of osteoarthritis patients is important for early diagnosis and treatment of osteoarthritis.

Objective To investigate the clinical efficacy of acupuncture plus Jisheng Shenqi pills in treating patients with diabetic neurogenic bladder. Method Patients with diabetic neurogenic bladder were allocated, using a random number table, to treatment and control groups. The treatment group received acupuncture plus Jisheng Shenqi pills and the control group, oral administration of Western medicine. Both groups of patients were treated for four weeks as a course. Post-treatment clinical total efficacy rates, pre-treatment and post-treatment postvoid residual urine volumes, and adverse reactions during treatment were compared between the two groups. Result The total efficacy rate was higher in the treatment group (93.8%) than in the control group (68.8%) (P<0.05). The postvoid residual urine volume decreased significantly in both groups of patients after treatment compared with before (P<0.05). Post-treatment postvoid residual urine volume was lower in the treatment group than in the control group (P<0.05). The adverse reaction rate was lower in the treatment group than in the control group (P<0.05). Conclusion Acupuncture plus Jisheng Shenqi pills is remarkably effective in treating patients with diabetic neurogenic bladder. It can effectively decrease postvoid residual urine volume in the patients, has higher safety and is worthy of clinical popularization.

Fusion PCR, which employs chimeric primers to generate PCR products with complementary ends in its amplifications, is a rapid and flexible method in joining different DNA fragments. This method can assemble DNA fragments without the treatment of restriction endonucleases and T4 DNA ligase. It offered a shortcut for the construction of homologous recombinant fragments. Through assembling three recombinant fragments successfully, fusion PCR procedure was improved and manipulation essentials of fusion PCR were described in detail. The results indicated that the improved procedure can assemble three or four fragments simultaneously, and the length of each fusion product is above 4.5 kb. The result recombinants were proposed to be use in further experiments, which structures had been confirmed by sequencing.

Based on a ?-tubulin gene EST sequence from Trichoderma harzianum mycelium cDNA library,a 1.74kb ?-tubulin gene coding region,together with a 1.5kb 5'-flanking region and a 1.0kb 3'-flanking region was cloned from T.harzianum by IPCR methods.The gene encodes a 446 amino acid polypeptide,which shows a high degree of homology with other fungal ?-tubulins.A three-dimensional model of the T.harzianum ?-tubulin was built based on the crystal structure of bovine tubulin using homology modeling method.Tertiary protein structure of the ?-tubulin should provide a basis for understanding a significant body of research on tubulin's properties and interactions with antimicrotubule agents.